Tetralin analogs having sphingosine 1-phosphate agonist activity

ABSTRACT

Tetralin analogs that have agonist activity at one or more of the S1P receptors are provided. The compounds are sphingosine analogs, which, after phosphorylation, can behave as agonists at S1P receptors.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application is a continuation of International applicationPCT/US2007/085392 filed on Nov. 21, 2007, and published in English as WO2008/064315 on May 29, 2008 which claims priority to ProvisionalApplication Nos. 60/956,376, filed Aug. 16, 2007 and Application Ser.No. 60/860,696, filed Nov. 21, 2006, the disclosures of all of which areincorporated by reference in their entirety.

US GOVERNMENT RIGHTS

This invention was made with United States Government support underGrant No. RO1 GM 067958 awarded by the National Institutes of Health.The United States Government has certain rights in the invention.

BACKGROUND OF THE INVENTION

Sphingosine 1-phosphate (S1P) is a lysophospholipid mediator that evokesa variety of cellular responses by stimulation of five members of theendothelial cell differentiation gene (EDG) receptor family. The EDGreceptors are G-protein coupled receptors (GPCRs) and on stimulationpropagate second messenger signals via activation of heterotrimericG-protein alpha (G_(α)) subunits and beta-gamma (G_(βγ)) dimers.Ultimately, this S1P-driven signaling results in cell survival,increased cell migration and, often, mitogenesis. The recent developmentof agonists targeting S1P receptors has provided insight regarding therole of this signaling system in physiologic homeostasis. For example,the immunomodulator, FTY720 (2-amino-2-[2-(4-octylphenyl) ethyl]propane1,3-diol), that following phosphorylation, is an agonist at 4 of 5 S1Preceptors, revealed that enhancing S1P tone influences lymphocytetrafficking. Further, S1P type 1 receptor (S1P₁) antagonists causeleakage of the lung capillary endothelium, which suggests that S1P maybe involved in maintaining the integrity of the endothelial barrier insome tissue beds.

Sphingosine 1-phosphate (S1P) is a lysophospholipid mediator that evokesa variety of cellular responses by stimulation of five members of theendothelial cell differentiation gene (EDG) receptor family.

Sphingosine-1-phosphate (S1P) has been demonstrated to induce manycellular processes, including those that result in platelet aggregation,cell proliferation, cell morphology, tumor-cell invasion, endothelialcell chemotaxis and angiogenesis. For these reasons, S1P receptors aregood targets for therapeutic applications such as wound healing andtumor growth inhibition.

Sphingosine-1-phosphate signals cells in part via a set of Gprotein-coupled receptors named S1P₁, S1P₂, S1P₃, S1P₄, and S1P₅(formerly EDG1, EDG5, EDG3, EDG6 and EDG8). The EDG receptors areG-protein coupled receptors (GPCRs) and on stimulation propagate secondmessenger signals via activation of heterotrimeric G-protein alpha(G_(α)) subunits and beta-gamma (G_(βγ)) dimers. These receptors share50-55% amino acid sequence identity and cluster with three otherreceptors (LPA₁, LPA₂, and LPA₃ (formerly EDG2, EDG4 and EDG7) for thestructurally related lysophosphatidic acid (LPA).

A conformational shift is induced in the G-Protein Coupled Receptor(GPCR) when the ligand binds to that receptor, causing GDP to bereplaced by GTP on the α-subunit of the associated G-proteins andsubsequent release of the G-proteins into the cytoplasm. The α-subunitthen dissociates from the βγ-subunit and each subunit can then associatewith effector proteins, which activate second messengers leading to acellular response. Eventually the GTP on the G-proteins is hydrolyzed toGDP and the subunits of the G-proteins reassociate with each other andthen with the receptor. Amplification plays a major role in the generalGPCR pathway. The binding of one ligand to one receptor leads to theactivation of many G-proteins, each capable of associating with manyeffector proteins leading to an amplified cellular response.

S1P receptors make good drug targets because individual receptors areboth tissue and response specific. Tissue specificity of the S1Preceptors is desirable because development of an agonist or antagonistselective for one receptor localizes the cellular response to tissuescontaining that receptor, limiting unwanted side effects. Responsespecificity of the S1P receptors is also of importance because it allowsfor the development of agonists or antagonists that initiate or suppresscertain cellular responses without affecting other responses. Forexample, the response specificity of the S1P receptors could allow foran S1P mimetic that initiates platelet aggregation without affectingcell morphology.

Sphingosine-1-phosphate is formed as a metabolite of sphingosine in itsreaction with sphingosine kinase and is stored in abundance in theaggregates of platelets where high levels of sphingosine kinase existand sphingosine lyase is lacking. S1P is released during plateletaggregation, accumulates in serum, and is also found in malignantascites. Reversible biodegradation of S1P most likely proceeds viahydrolysis by ectophosphohydrolases, specifically the sphingosine1-phosphate phosphohydrolases. Irreversible degradation of S1P iscatalyzed by S1P lyase yielding ethanolamine phosphate and hexadecena

Currently, there is a need for novel, potent, and selective agents thatare agonists of the S1P receptor having enhanced potency, selectivity,and oral bioavailability. In addition, there is a need in the art foridentification of, as well as the synthesis and use of such compounds.The present invention satisfies these needs.

SUMMARY

The present invention provides in one aspect compounds that have agonistactivity at one or more of the S1P receptors. The compounds aresphingosine analogs that, after phosphorylation, can behave as agonistsat S1P receptors. Accordingly, there is provided compounds of Formula I:

-   -   wherein X¹, Y¹ and Z¹ are independently O, CR^(a), CR^(a)R^(b),        N, NR^(c), or S.    -   R¹ and R² are independently hydrogen, halo, halo(C₁-C₁₀)alkyl,        cyano, —NR^(a)R^(b), (C₁-C₂₀)alkyl, (C₂-C₂₀)alkenyl,        (C₂-C₂₀)alkynyl, (C₁-C₂₀)alkoxy, (C₂-C₂₆)alkoxyalkyl,        (C₃-C₁₂)cycloalkyl, (C₆-C₁₀)aryl, (C₇-C₃₀)arylalkyl,        (C₂-C₁₀)heterocyclic, (C₄-C₁₀)heteroaryl, or        (C₄-C₁₀)heteroaryl(C₁-C₂₀)alkyl; or    -   R² can be a group having Formula II, III, IV, V, or VI:

-   -   wherein each R⁷, R⁸, R⁹, R¹⁰, R¹¹, R¹², R¹³, and R¹⁴ is        independently O, S, C, CR¹⁵, CR¹⁶R¹⁷, C═O, N or NR¹⁸; and    -   each R¹⁵, R¹⁶ and R¹⁷ is independently hydrogen, halo,        (C₁-C₁₀)alkyl, (C₆-C₁₀)aryl, (C₁-C₁₀)alkyl substituted with        halo, hydroxy, (C₁-C₁₀)alkoxy, or cyano; and where R¹⁸ can be        hydrogen or (C₁-C₁₀)alkyl;    -   Z² is hydrogen, halo, halo(C₁-C₁₀)alkyl, cyano, —NR^(a)R^(b),        (C₁-C₂₀)alkyl, (C₂-C₂₀)alkenyl, (C₂-C₂₀)alkynyl, (C₁-C₂₀)alkoxy,        (C₂-C₂₆)alkoxyalkyl, (C₃-C₁₂)cycloalkyl, (C₆-C₁₀)aryl,        (C₇-C₃₀)arylalkyl, (C₂-C₁₀)heterocyclic, (C₄-C₁₀)heteroaryl, or        (C₄-C₁₀)heteroaryl(C₁-C₂₀)alkyl. The alkyl, alkenyl, alkynyl,        cycloalkyl, aryl, heterocyclic, or heteroaryl groups of Z² are        optionally perfluorinated or optionally substituted with 1, 2,        3, or 4 groups where the substituent groups are independently        hydroxy, halo, cyano, (C₁-C₁₀)alkoxy, C₆-aryl,        (C₇-C₂₄)arylalkyl, oxo (═O), or imino (═NR^(d)), wherein one or        more of the carbon atoms in the Z² alkyl groups can be        independently replaced with non-peroxide oxygen, sulfur or        NR^(c);

-   -   indicates one or more optional double bonds;    -   Y² is a bond (absent), O, S, C═O, or NR^(c), CH₂; W¹ is a bond        (absent); —CH₂— and m is 1, 2, or 3, or (C═O)(CH₂)₁₋₅ and m is        1; wherein W¹ is optionally interrupted with non-peroxide O, S,        C═O, or NR^(c). Each         represents an optional double bond; and n is 0, 1, 2, or 3.

R³ is hydrogen, (C₁-C₁₀)alkyl, hydroxy(C₁-C₁₀)alkyl or (C₁-C₁₀)alkoxy;and R⁴ is hydroxy (—OH), phosphate (—OPO₃H₂), phosphonate (—CH₂PO₃H₂),or alpha-substituted phosphonate. Each R^(a), R^(b), R^(c), and Rd isindependently hydrogen, or (C₁-C₁₀)alkyl.

The alkyl, alkenyl, alkynyl, cycloalkyl, aryl, heterocyclic, orheteroaryl groups of R¹ and R² independently are optionallyperfluorinated or optionally substituted with 1, 2, 3, or 4 groups wherethe substituent groups are independently hydroxy, halo, cyano,(C₁-C₁₀)alkoxy, C₆-aryl, (C₇-C₂₄)arylalkyl, oxo (═O), or imino(═NR_(d)), wherein one or more of the carbon atoms in the R¹ or R² alkylgroups can be independently replaced with non-peroxide oxygen, sulfur orNR^(c). The alkyl groups of R³ are optionally substituted with 1, or 2hydroxy groups; and Rd is hydrogen, or (C₁-C₁₀)alkyl. The inventionincludes pharmaceutically acceptable salts or esters of the compounds ofFormula I.

In another aspect, the invention provides phosphate monoesters havingFormula VIII.

where R¹, R², R³, X¹, Y¹, and Z¹ are as defined above. In anotheraspect, the invention provides enantiomers and stereoisomers of thecompounds having Formulas I, or VIII.

In another aspect, the invention provides pro-drugs of the compounds ofFormula I. The invention also provides compounds of Formula I orpharmaceutically acceptable salts or esters thereof for use in medicaltherapy.

In another aspect, the present invention provides a method forinhibiting angiogenesis in a tumor, comprising contacting the cancerouscells with an effective amount of a compound of Formula I, or apharmaceutically acceptable salt thereof.

In another aspect, the invention provides a method for modulating theimmune system by altering lymphocyte trafficking for treatment ofautoimmune diseases or prolongation of allograft transplant survival,said method comprising administering an effective amount of at least onecompound of the invention to a subject in need thereof.

In another aspect, the invention provides a method for treatingneuropathic pain, wherein a pharmaceutical composition comprising aneffective amount of at least one compound of the invention and apharmaceutically-acceptable carrier is administered to a subject in needthereof.

In another aspect, the invention provides a method for repairingvascular injury following catheterization, comprising contacting thelumen of the affected vessel with an effective amount of the compound ofFormula I. In another aspect, the invention includes coating indwellingstents with a compound of Formula I.

In another aspect, the invention provides a compound of Formula I, or apharmaceutically acceptable salt thereof for use in medical treatment(for example, treatment of neoplastic disease, treatment of neuropathicpain, treatment of autoimmune disease, prolongation of allograftsurvival).

In another aspect, the invention provides a method for the use of acompound of Formula I or a pharmaceutically acceptable salt thereof toprepare a medicament for inhibiting tumor growth, metastasis or tumorangiogenesis in a mammalian species (for example, a human).

In another aspect, the invention provides for the use of a compound ofFormula I or a pharmaceutically acceptable salt thereof to prepare amedicament for treating an autoimmune disease or prolonging allograftsurvival in a mammalian species (for example, a human).

In another aspect, the invention provides for the use of a compound ofFormula I or a pharmaceutically acceptable salt thereof to prepare amedicament for treating neuropathic pain in a mammalian species (forexample, a human).

In another aspect, the invention provides a method for assessing acompound of Formula I (e.g., S1P receptor pro-drugs) as a substrate forsphingosine kinase types 1 or 2, in vitro and in vivo. The inventionincludes also a method of assessing a compound of Formula (II) forbinding to designated receptor sites comprising in vivo or in vitro,with an amount of a compound of Formula (II) effective to bind saidreceptors. Tissue comprising ligand bound designated S1P receptor sitescan be used to measure the selectivity of test compounds for specificreceptor subtypes, or can be used as a tool to identify potentialtherapeutic agents for the treatment of diseases, by contacting saidagents with said ligand-receptor complexes, and measuring the extent ofdisplacement of the ligand and/or binding of the agent.

In another aspect, the invention provides novel intermediates andprocesses disclosed herein that are useful for preparing compounds ofFormula I, including the generic and specific intermediates as well asthe synthetic processes described herein.

In another aspect, the present invention provides synthetic schemes andmethods of use of compounds having Formula I and analogs or derivativesthereof. The invention further provides synthetic and modificationschemes for preparing analogs and derivatives of the compounds of theinvention, as well as compositions and methods for the use of suchanalogs and derivatives.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 illustrate three S1P agonists FTY720, AAL151 and compound XXIX.

FIGS. 2A-2C illustrate additional compounds of Formula I.

FIGS. 3-5 illustrate syntheses of compounds of Formula I.

DETAILED DESCRIPTION

The following abbreviations are used herein: S1P,sphingosine-1-phosphate; S1P₁₋₅-S1P receptor types; GPCR, G-proteincoupled receptor; SAR, structure-activity relationship; EDG, endothelialcell differentiation gene; EAE, experimental autoimmuneencephalomyelitis; NOD non-obese diabetic; TNFα, tumor necrosis factoralpha; HDL, high density lipoprotein; and RT-PCR, reverse transcriptasepolymerase chain reaction.

In describing and claiming the invention, the following terminology willbe used in accordance with the definitions set forth below. Unlessotherwise defined, all technical and scientific terms used herein havethe same meaning as commonly understood by one of ordinary skill in theart to which this invention belongs. Although any materials and methodssimilar or equivalent to those described herein can be used in thepractice or testing of the present invention, the preferred materialsand methods are described herein. As used herein, each of the followingterms has meaning associated with it in this section. Specific andpreferred values listed below for radicals, substituents, and ranges arefor illustrations only; they do not exclude other defined values orother values within defined ranges for the radicals and substituents.

The terms “a,” “an,” “the,” “at least one,” and “one or more” are usedinterchangeably. Thus, for example, a composition that comprises “an”element means one element or more than one element.

The term “receptor agonists” are compounds that mimic the action of S1Pat one or more of its receptors but may have differing potency and/orefficacy.

The term “receptor antagonists” are compounds that 1) lack intrinsicagonist activity and 2) block agonist (e.g., S1P) activation of the S1Preceptor(s), often in a manner that is both fully surmountable andreversible (‘competitive antagonist’).

The term “affected cell” refers to a cell of a subject afflicted with adisease or disorder, which affected cell has an altered phenotyperelative to a subject not afflicted with a disease or disorder.

Cells or tissue are “affected” by a disease or disorder if the cells ortissue have an altered phenotype relative to the same cells or tissue ina subject not afflicted with a disease or disorder.

A disease or disorder is “alleviated” if the severity of a symptom ofthe disease or disorder, the frequency with which such a symptom isexperienced by a patient, or both, is reduced.

An “analog” of a chemical compound is a compound that, by way ofexample, resembles another in structure but is not necessarily an isomer(e.g., 5-fluorouracil is an analog of thymine).

The terms “cell,” “cell line,” and “cell culture” may be usedinterchangeably.

A “control” cell, tissue, sample, or subject is a cell, tissue, sample,or subject of the same type as a test cell, tissue, sample, or subject.The control may, for example, be examined at precisely or nearly thesame time the test cell, tissue, sample, or subject is examined. Thecontrol may also, for example, be examined at a time distant from thetime at which the test cell, tissue, sample, or subject is examined, andthe results of the examination of the control may be recorded so thatthe recorded results may be compared with results obtained byexamination of a test cell, tissue, sample, or subject. The control mayalso be obtained from another source or similar source other than thetest group or a test subject, where the test sample is obtained from asubject suspected of having a disease or disorder for which the test isbeing performed.

A “test” cell, tissue, sample, or subject is one being examined ortreated.

A “pathoindicative” cell, tissue, or sample is one which, when present,is an indication that the animal in which the cell, tissue, or sample islocated (or from which the tissue was obtained) is afflicted with adisease or disorder. By way of example, the presence of one or morebreast cells in a lung tissue of an animal is an indication that theanimal is afflicted with metastatic breast cancer.

A tissue “normally comprises” a cell if one or more of the cell arepresent in the tissue in an animal not afflicted with a disease ordisorder.

The use of the word “detect” and its grammatical variants is meant torefer to measurement of the species without quantification, whereas useof the word “determine” or “measure” with their grammatical variants aremeant to refer to measurement of the species with quantification. Theterms “detect” and “identify” are used interchangeably herein.

A “detectable marker” or a “reporter molecule” is an atom or a moleculethat permits the specific detection of a compound comprising the markerin the presence of similar compounds without a marker. Detectablemarkers or reporter molecules include, e.g., radioactive isotopes,antigenic determinants, enzymes, nucleic acids available forhybridization, chromophores, fluorophores, chemiluminescent molecules,electrochemically detectable molecules, and molecules that provide foraltered fluorescence-polarization or altered light-scattering.

A “disease” is a state of health of an animal wherein the animal cannotmaintain homeostasis, and wherein if the disease is not ameliorated thenthe animal's health continues to deteriorate.

A “disorder” in an animal is a state of health in which the animal isable to maintain homeostasis, but in which the animal's state of healthis less favorable than it would be in the absence of the disorder. Leftuntreated, a disorder does not necessarily cause a further decrease inthe animal's state of health.

An “effective amount” means an amount sufficient to produce a selectedeffect. For example, an effective amount of an S1P receptor antagonistis an amount that decreases the cell signaling activity of the S1Preceptor.

A “functional” molecule is a molecule in a form in which it exhibits aproperty by which it is characterized. By way of example, a functionalenzyme is one which exhibits the characteristic catalytic activity bywhich the enzyme is characterized.

The term “inhibit” refers to the ability of a compound of the inventionto reduce or impede a described function. Preferably, inhibition is byat least 10%, more preferably by at least 25%, even more preferably byat least 50%, and most preferably, the function is inhibited by at least75%.

“Instructional material” includes a publication, a recording, a diagram,or any other medium of expression which can be used to communicate theusefulness of the peptide of the invention in the kit for effectingalleviation of the various diseases or disorders recited herein.Optionally, or alternately, the instructional material may describe oneor more methods of alleviating the diseases or disorders in a cell or atissue of a mammal. The instructional material of the kit of theinvention may, for example, be affixed to a container which contains theidentified compound invention or be shipped together with a containerwhich contains the identified compound. Alternatively, the instructionalmaterial may be shipped separately from the container with the intentionthat the instructional material and the compound be used cooperativelyby the recipient.

The term “parenteral” means not through the alimentary canal but by someother route such as subcutaneous, intramuscular, intraspinal, orintravenous.

The term “pharmaceutically acceptable carrier” includes any of thestandard pharmaceutical carriers, such as a phosphate buffered salinesolution, water and emulsions such as an oil/water or water/oilemulsion, and various types of wetting agents. The term also encompassesany of the agents approved by a regulatory agency of the U.S. Federalgovernment or listed in the U.S. Pharmacopeia for use in animals,including humans.

The term “purified” and similar terms relate to the isolation of amolecule or compound in a form that is substantially free (at least 75%free, preferably 90% free, and most preferably at least 95% free) fromother components normally associated with the molecule or compound in anative environment. The term “purified” does not necessarily indicatethat complete purity of the particular molecules achieved during theprocess. A “very pure” compound refers to a compound that is greaterthan 90% pure. A “highly purified” compound refers to a compound that isgreater than 95% pure.

A “sample” refers preferably to a biological sample from a subject,including, but not limited to, normal tissue samples, diseased tissuesamples, biopsies, blood, saliva, feces, semen, tears, and urine. Asample can also be any other source of material obtained from a subject,which contains cells, tissues, or fluid of interest. A sample can alsobe obtained from cell or tissue culture.

The term “standard,” refers to something used for comparison. Forexample, a standard can be a known standard agent or compound which isadministered or added to a control sample and used for comparing resultswhen measuring said compound in a test sample. Standard can also referto an “internal standard,” such as an agent or compound which is addedat known amounts to a sample and is useful in determining such things aspurification or recovery rates when a sample is processed or subjectedto purification or extraction procedures before a marker of interest ismeasured.

A “subject” of analysis, diagnosis, or treatment is an animal. Suchanimals include mammals, preferably a human.

A “therapeutic” treatment is a treatment administered to a subject whoexhibits signs of pathology for the purpose of diminishing oreliminating those signs.

A “therapeutically effective amount” of a compound is that amount ofcompound which is sufficient to provide a beneficial effect to thesubject to which the compound is administered.

The term “treating” includes prophylaxis of the specific disorder orcondition, or alleviation of the symptoms associated with a specificdisorder or condition and/or preventing or eliminating said symptoms.

The disclosed compounds are generally named according to the IUPAC orCAS nomenclature system. Abbreviations which are well known to one ofordinary skill in the art may be used (e.g., “Ph” for phenyl, “Me” formethyl, “Et” for ethyl, “h” for hour or hours, “rt” for roomtemperature, “THF” for tetrahydrofuran, and “rac” for racemic mixture).

Specific and preferred values listed below for radicals, substituents,and ranges, are for illustration only; they do not exclude other definedvalues or other values within defined ranges for the radicals andsubstituents. The compounds of the invention include compounds ofFormula I having any combination of the values, specific values, morespecific values, and preferred values described herein.

Specifically, the term “halogen” or “halo” includes bromo, chloro,fluoro, and iodo. The term “haloalkyl”, refers to an alkyl radicalbearing at least one halogen substituent, non-limiting examples include,but are not limited to, chloromethyl, fluoroethyl or trifluoromethyl andthe like. The term “C₁-C₂₀ alkyl” refers to a branched or linear alkylgroup having from one to twenty carbons. Non-limiting examples include,but are not limited to, methyl, ethyl, n-propyl, iso-propyl, butyl,iso-butyl, sec-butyl, tert-butyl, pentyl, hexyl, heptyl, octyl and thelike. The term “C₂-C₂₀ alkenyl”, refers to an olefinically unsaturatedbranched or linear group having from two to twenty carbon atoms and atleast one double bond. Typically, C₂-C₂₀ alkenyl groups include, but arenot limited to, 1-propenyl, 2-propenyl, 1,3-butadienyl, 1-butenyl,hexenyl, pentenyl, hexenyl, heptenyl, octenyl and the like. The term(C₂-C₂₀)alkynyl can be ethynyl, 1-propynyl, 2-propynyl, 1-butynyl,2-butynyl, 3-butynyl, 1-pentynyl, 2-pentynyl, 3-pentynyl, 4-pentynyl,1-hexynyl, 2-hexynyl, 3-hexynyl, 4-hexynyl, or 5-hexynyl, and the like.The term “(C₁-C₂₀)alkoxy or alkoxyl” refers to an alkyl group attachedthrough an oxygen atom. Examples of (C₁-C₂₀)alkoxy can be methoxy,ethoxy, propoxy, isopropoxy, butoxy, iso-butoxy, sec-butoxy, pentoxy,3-pentoxy, hexyloxy, heptoxy, or octoxy and the like.

The term “C₃-C₁₂ cycloalkyl”, can be cyclopropyl, cyclobutyl,cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl and the like.

The term “optionally substituted” refers to from zero, one, two, threeor four substituents, wherein the substituents are each independentlyselected. Each of the independently selected substituents may be thesame or different than other substituents.

The term “(C₆-C₁₀)aryl” refers to a mono or bicyclic carbocyclic ringsystem having one or two aromatic rings including, but not limited to,phenyl, benzyl, naphthyl, tetrahydronaphthyl, indanyl, indenyl, and thelike. The term “aryl” refers to a mono or bicyclic carbocyclic ringsystem having one or two aromatic rings including, but not limited to,phenyl, benzyl, naphthyl, tetrahydronaphthyl, indanyl, indenyl, and thelike.

The term “aryl(C₁-C₂₀)alkyl” or “aralkyl” refers to an alkyl groupsubstituted with a mono or bicyclic carbocyclic ring system having oneor two aromatic rings including, a group such as phenyl, naphthyl,tetrahydronaphthyl, indanyl, indenyl, and the like. Non-limitingexamples of arylalkyl include benzyl, phenylethyl, and the like.

The term “optionally substituted aryl” includes aryl compounds havingfrom zero to four substituents, and a substituted aryl includes arylcompounds having one to three substituents, wherein the substituentsinclude groups such as, for example, alkyl, halo, or amino substituents.

The “(C₂-C₁₀)heterocyclic group” refers to an optionally substitutedmono- or bicyclic carbocyclic ring system containing one, two, or threeheteroatoms (optionally in each ring) wherein the heteroatoms areoxygen, sulfur, and nitrogen.

The term “bicyclic” represents either an unsaturated or saturated stable7- to 12-membered bridged or fused bicyclic carbon ring. The bicyclicring may be attached at any carbon atom which affords a stablestructure. The term includes, but is not limited to, naphthyl,dicyclohexyl, dicyclohexenyl, and the like.

The term “phosphate analog” and “phosphonate analog” comprise analogs ofphosphate and phosphonate wherein the phosphorous atom is in the +5oxidation state and one or more of the oxygen atoms is replaced with anon-oxygen moiety, including for example, the phosphate analogsphosphorothioate, phosphorodithioate, phosphoroselenoate,phosphorodiselenoate, phosphoroanilothioate, phosphoranilidate,phosphoramidate, boronophosphates, and the like, including associatedcounterions, e.g., H, NH₄, Na, K, and the like if such counterions arepresent.

The term “alpha-substituted phosphonate” includes phosphonate(—CH₂PO₃H₂) groups that are substituted on the alpha-carbon such as—CHFPO₃H₂, —CF₂PO₃H₂, —CHOHPO₃H₂, —C═OPO₃H₂) and the like.

A “derivative” of a compound refers to a chemical compound that may beproduced from another compound of similar structure in one or moresteps, as in replacement of hydrogen by an alkyl, acyl, or amino group.

The term “pharmaceutically acceptable carrier” includes any of thestandard pharmaceutical carriers, such as a phosphate buffered salinesolution, hydroxypropyl beta-cyclodextrins (HO-propyl betacyclodextrins), water, emulsions such as an oil/water or water/oilemulsion, and various types of wetting agents. The term also encompassesany of the agents approved by a regulatory agency of the US Federalgovernment or listed in the US Pharmacopeia for use in animals,including humans.

The term “pharmaceutically-acceptable salt” refers to salts which retainthe biological effectiveness and properties of the compounds of thepresent invention and which are not biologically or otherwiseundesirable. In many cases, the compounds of the present invention arecapable of forming acid and/or base salts by virtue of the presence ofamino and/or carboxyl groups or groups similar thereto.

An “effective amount” means an amount sufficient to produce a selectedeffect. For example, an effective amount of an S1P receptor agonist isan amount that decreases the cell signaling activity of the S1Preceptor.

The disclosed compounds can contain one or more asymmetric centers inthe molecule. In accordance with the present disclosure any structurethat does not designate the stereochemistry is to be understood asembracing all the various optical isomers, as well as racemic mixturesthereof.

The disclosed compounds can contain one or more asymmetric centers inthe molecule. In accordance with the present disclosure any structurethat does not designate the stereochemistry is to be understood asembracing all the various optical isomers, as well as racemic mixturesthereof.

The compounds of the present invention may exist in tautomeric forms andthe invention includes both mixtures and separate individual tautomers.For example, the following structure:

is understood to represent a mixture of the structures:

The terms 16:0, 18:0, 18:1, 20:4 or 22:6 hydrocarbon refers to abranched or straight alkyl or alkenyl group, wherein the first integerrepresents the total number of carbons in the group and the secondinteger represent the number of double bonds in the group.

An “S1P modulating agent” refers a compound or composition that iscapable of inducing a detectable change in S1P receptor activity in vivoor in vitro (e.g., at least 10% increase or decrease in S1P activity asmeasured by a given assay such as the bioassay described in the examplesand known in the art. “S1P receptor,” as used herein, refers to all ofthe S1P receptor subtypes (for example, the S1P receptors S1P₁, S1P₂,S1P₃, S1P₄, and S1P₅), unless the specific subtype is indicated.

The disclosed compounds can contain one or more asymmetric centers inthe molecule. In accordance with the present disclosure any structurethat does not designate the stereochemistry is to be understood asembracing all the various optical isomers, as well as racemic mixturesthereof.

It will be appreciated by those skilled in the art that compounds of theinvention having chiral centers may exist in and be isolated inoptically active and racemic forms. It is to be understood that thepresent invention encompasses any racemic, optically active orstereoisomeric form, or mixtures thereof, of a compound of theinvention, which possess the useful properties described herein, such asthe S,R; S,S; R,R; or R,S diastereomers. It is well known in the art howto prepare such optically active forms (for example, by resolution ofthe racemic form by recrystallization techniques, by synthesis fromoptically-active starting materials, by chiral synthesis, or bychromatographic separation using a chiral stationary phase) and how todetermine S1P agonist activity using the standard tests describedherein, or using other similar tests which are well known in the art. Inaddition, some compounds may exhibit polymorphism.

Potential uses of an S1P receptor agonist pro-drugs (S1P₁ receptor typeselective agonists preferred) include, but are not limited to, alteringlymphocyte trafficking as a method of treatment for autoimmunepathologies such as uveitis, type I diabetes, rheumatoid arthritis,inflammatory bowel diseases, and, most particularly, multiple sclerosis.“Treatment” of multiple sclerosis includes the various forms of thedisease including relapsing-remitting, chronic progressive, etc., andthe S1P receptor agonists can be used alone or in conjunction with otheragents to relieve signs and symptoms of the disease as well asprophylactically.

In addition, the disclosed compounds of the invention can be used foraltering lymphocyte trafficking is a method for prolonging allograftsurvival, for example solid organ transplants, treatment of graft vs.host disease, bone marrow transplantation, and the like.

In addition, the disclosed compounds of the invention can be used toinhibit autotaxin. Autotaxin, a plasma phosphodiesterase, has beendemonstrated to undergo end product inhibition. Autotaxin hydrolyzesseveral substrates to yield lysophosphatidic acid and sphingosine1-phosphate, and has been implicated in cancer progression andangiogenesis. Therefore, S1P receptor agonist pro-drugs of the inventioncan be used to inhibit autotaxin. This activity may be combined withagonism at S1P receptors or may be independent of such activity.

In addition, disclosed compounds of the invention can be useful forinhibition of S1P lyase. S1P lyase is an intracellular enzyme thatirreversibly degrades S1P. Inhibition of S1P lyase disrupts lymphocytetrafficking with concomitant lymphopenia. Accordingly, S1P lyaseinhibitors can be useful in modulating immune system function.Therefore, the compounds of the invention can be used to inhibit S1Plyase. This inhibition could be in concert with S1P receptor activity,or be independent of activity at any S1P receptor.

In addition, disclosed compounds of the invention can be useful asantagonists of the cannabinoid CB₁ receptor. CB₁ antagonism isassociated with a decrease in body weight and an improvement in bloodlipid profiles. The CB₁ antagonism could be in concert with S1P receptoractivity, or be independent of activity at any S1P receptor.

In addition, disclosed compounds of the invention can be useful forinhibition of group IVA cytosolic PLA₂ (cPLA₂). cPLA₂ catalyzes therelease of eicosanoic acids (e.g. arachidonic acid). The eicosanoicacids are transformed to pro-inflammatory eicosanoids such asprostaglandins and leukotrienes. Thus compounds of the invention can beuseful as anti-inflammatory agents. This inhibition could be in concertwith S1P receptor activity, or be independent of activity at any S1Preceptor.

In addition, disclosed compounds of the invention can be useful forinhibition of the multiple substrate lipid kinase (MuLK). MuLK is highlyexpressed in many human tumor cells and thus its inhibition might slowthe growth or spread of tumors.

“Treatment” of multiple sclerosis includes the various forms of thedisease including relapsing-remitting, chronic progressive, etc., andthe S1P receptor agonists can be used alone or in conjunction with otheragents to relieve signs and symptoms of the disease as well asprophylactically.

In another aspect, the present invention provides compositions andmethods for the use of the S1P analogs of the invention to prevent,inhibit, or treat neuropathic pain by agonizing or antagonizing the S1Preceptors. Pain can be nociceptive or neuropathic in nature. Neuropathicpain is characterized by its chronic nature, an absence of an obvious,direct cause (i.e., tissue damage), and allodynia. Allodynia is theperception of normal stimuli as painful (examples include the touch ofclothing, warm or cool air, etc.). Neuropathic pain is often a sequel tonerve damage in an extremity such as an arm, or more often, a leg.Typically, neuropathic pain is not responsive to opiates ornon-steroidal anti-inflammatory drugs such as aspirin.

In another aspect, the present invention provides compositions andmethods for the use of S1P analogs to prevent and inhibit vascularrestenosis following vascular injury. In one aspect, the injury can bedue to balloon angioplasty. The present invention further providesmethods for treating subjects to prevent vascular restenosis.

In another aspect, the present invention provides compositions andmethods for the use of sphingosine analogs (including S1P pro-drugs) toprevent asthma attacks. In one aspect, the asthma could be due to overproduction of cysteinyl leukotrienes. The present invention furtherprovides methods for treating subjects to treat asthma.

In another aspect, the present invention provides compositions andmethods for the use of sphingosine analogs (including S1P pro-drugs) totreat obesity.

In another aspect, the present invention provides compositions andmethods for the use of sphingosine analogs (including S1P pro-drugs) tonormalize blood lipid composition. In one aspect, blood low densitylipoprotein (LDL or ‘bad cholesterol’) levels could be lowered. Inanother aspect, blood triglyceride levels could be lowered.

In another aspect, the present invention provides compositions andmethods for the use of S1P analogs and S1P pro-drugs for the preventionand treatment of arteriosclerosis.

In another aspect, the present invention provides compositions andmethods for the use of S1P analogs and S1P pro-drugs for the treatmentof neoplastic disease. In one aspect, this treatment is effected byapplication of S1P receptor antagonists that are efficacious by virtueof their anti-angiogenic properties. In another aspect, the treatment iseffected by administration of sphingosine analogs that inhibit themultiple substrate lipid kinase.

In another aspect, the present invention provides compositions andmethods for the use of S1P analogs and S1P pro-drugs for the treatmentof neurodegenerative diseases. In one aspect, the treatment is forsenile dementia of the Alzheimers type.

The present invention is also includes pharmaceutical compositionscomprising the disclosed compounds of the present invention. Moreparticularly, such compounds can be Formulated as pharmaceuticalcompositions using standard pharmaceutically acceptable carriers,fillers, solubilizing agents and stabilizers known to those skilled inthe art. For example, a pharmaceutical composition including a disclosedcompound, analog, derivative, or modification thereof, as describedherein, is used to administer the appropriate compound to a subject.

The compounds of the invention are useful for treating a disease ordisorder including administering to a subject in need thereof of atherapeutically acceptable amount of a compound of Formula (I), or apharmaceutical composition comprising a therapeutically effective amountof a compound of Formula (I), and a pharmaceutically-acceptable carrier.

The disclosed compounds and method are directed to sphingosine1-phosphate (S1P) analogs that have activity as receptor receptoragonists or antagonists at one or more S1P receptors, specifically theS1P₁, S1P₄ and S1P₅ receptor types. The disclosed compound and methodinclude both compounds that have a phosphate moiety as well as compoundswith hydrolysis-resistant phosphate surrogates such as phosphonates,alpha-substituted phosphonates particularly where the alpha substitutionis a halogen and phosphothionates.

Specific and preferred values listed below for radicals, substituents,and ranges, are for illustration only; they do not exclude other definedvalues or other values within defined ranges for the radicals andsubstituents.

Exemplary values for X¹, Y¹ and Z¹ are independently O, CH, CH₂, CHCF₃,N, NH, or S.

An additional exemplary value for X¹, Y¹ and Z¹ is CH₂.

An exemplary compound has the Formula

Exemplary values for R¹ include hydrogen, fluorine, chlorine, bromine,trifluoro-methyl, methoxy, (C₁-C₆)alkyl, (C₁-C₆)haloalkyl, or(C₁-C₆)alkyl substituted with, alkoxy or cyano.

More values for R¹ are hydrogen, trifluoro-methyl, or —CH₂CF₃.

Additional exemplary values for R¹ are alkyl-substituted aryl,aryl-substituted alkyl, or aryl-substituted arylalkyl.

Even more exemplary values for R¹ are benzyl, phenylethyl, or benzylsubstituted with methyl.

Exemplary values for R² include

An exemplary value for W¹ is a bond, —CH₂—CH₂—CH₂— or —(C═O)(CH₂)₁₋₅.

Additional for R² having Formula VI are

-   -   wherein Z² is (CH₃)₃C—, CH₃CH₂(CH₃)₂C—, CH₃CH₂CH₂—,        CH₃(CH₂)₂CH₂—, CH₃(CH₂)₄CH₂—, (CH₃)₂CHCH₂—, (CH₃)₃CCH₂—,        CH₃CH₂O—, (CH₃)₂CHO—, or CF₃CH₂CH₂— or a group having the        formula:

An additional value for an R² groups having Formula VI (para substituted3,5-diphenyl-(1,2,4)-oxadiazoles) is;

Another value for R² groups having Formula VI is;

Another specific value for R² having Formula II is;

Another exemplary value for an R² group having Formula II is;

Additional values for R² groups having Formula III are;

Another value for R² having formula V is;

Additional exemplary values for R² include (C₁-C₂₀)alkyl,(C₁-C₂₀)alkoxy, or (C₂-C₂₆)alkoxyalkyl.

Additional exemplary values for R² include (C₁-C₂₀)alkyl,(C₁-C₂₀)alkoxy, or (C₂-C₂₆)alkoxyalkyl.

More exemplary values for R² include (C₁-C₁₀)alkyl, (C₂-C₁₀)alkenyl and(C₂-C₁₀)alkynyl or (C₁-C₁₀)alkoxy optionally substituted with carbonyl(C═O) or oxime (C═NR^(d)) groups.

Additional values for R² include methyl, ethyl, propyl, butyl, pentyl,hexyl, heptyl, octyl, trifluoromethyl, trifluoroethyl, trifluoromethoxy,trifluoroethoxy, methoxy, ethoxy, propoxy, butoxy, pentoxy, heptoxy, oroctoxy.

Exemplary values for R³ include methyl, hydroxymethyl, ethyl,hydroxyethyl, propyl, hydroxypropyl, or isopropyl.

Additional values for R³ are methyl, hydroxymethyl, ethyl, orhydroxyethyl.

Exemplary values for R⁴ include is hydroxy, or phosphate (—OPO₃H₂).

ADDITIONAL EMBODIMENTS

Additional embodiments of the invention include:

-   1. A compound of Formula I:

-   -   wherein    -   X¹, Y¹ and Z¹ are independently O, CR^(a), CR^(a)R^(b), N,        NR^(c), or S;    -   R¹ and R² are independently hydrogen, halo, halo(C₁-C₁₀)alkyl,        cyano, —NR^(a)R^(b), (C₁-C₂₀)alkyl, (C₂-C₂₀)alkenyl,        (C₂-C₂₀)alkynyl, (C₁-C₂₀)alkoxy, (C₂-C₂₆)alkoxyalkyl,        (C₃-C₁₂)cycloalkyl, (C₆-C₁₀)aryl, (C₇-C₃₀)arylalkyl,        (C₂-C₁₀)heterocyclic, (C₄-C₁₀)heteroaryl, or        (C₄-C₁₀)heteroaryl(C₁-C₂₀)alkyl;    -   wherein the alkyl, alkenyl, alkynyl, cycloalkyl, aryl,        heterocyclic, or heteroaryl groups of R¹ and R² independently        are optionally perfluorinated or optionally substituted with 1,        2, 3, or 4 groups where the substituent groups are independently        hydroxy, halo, cyano, (C₁-C₁₀)alkoxy, C₆-aryl,        (C₇-C₂₄)arylalkyl, oxo (═O), or imino (═NR^(d)), wherein one or        more of the carbon atoms in the R¹ or R² alkyl groups can be        independently replaced with non-peroxide oxygen, sulfur or        NR^(c); the alkyl groups of R³ are optionally substituted with        1, or 2 hydroxy groups; or    -   R² can be a group having formula II, III, IV, V, or VI;

-   -   wherein each R⁷, R⁸, R⁹, R¹⁰, R¹¹, R¹², R¹³, and R¹⁴ is        independently O, S, C, CR¹⁵, CR¹⁶R¹⁷, C═O, N or NR¹⁸;    -   each R¹⁵, R¹⁶ and R¹⁷ is independently hydrogen, halo,        (C₁-C₁₀)alkyl, (C₆-C₁₀)aryl, (C₁-C₁₀)alkyl substituted with        halo, hydroxy, (C₁-C₁₀)alkoxy, or cyano; and where R¹⁸ can be        hydrogen or (C₁-C₁₀)alkyl;    -   where Z² is hydrogen, halo, halo(C₁-C₁₀)alkyl, cyano,        —NR^(a)R^(b), (C₁-C₂₀)alkyl, (C₂-C₂₀)alkenyl, (C₂-C₂₀)alkynyl,        (C₁-C₂₀)alkoxy, (C₂-C₂₆)alkoxyalkyl, (C₃-C₁₂)cycloalkyl,        (C₆-C₁₀)aryl, (C₇-C₃₀)arylalkyl, (C₂-C₁₀)heterocyclic,        (C₄-C₁₀)heteroaryl, or (C₄-C₁₀)heteroaryl(C₁-C₂₀)alkyl; wherein        the alkyl, alkenyl, alkynyl, cycloalkyl, aryl, heterocyclic, or        heteroaryl groups of Z² are optionally perfluorinated or        optionally substituted with 1, 2, 3, or 4 groups where the        substituent groups are independently hydroxy, halo, cyano,        (C₁-C₁₀)alkoxy, C₆-aryl, (C₇-C₂₄)arylalkyl, oxo (═O), or imino        (═NR^(d)), wherein one or more of the carbon atoms in the Z²        alkyl groups can be independently replaced with non-peroxide        oxygen, sulfur or NR^(c);

-   -   indicates one or more optional double bonds;    -   wherein Y² is a bond, O, S, C═O, or NR^(c), CH₂; W¹ is a bond,        —CH₂— and m is 1, 2, or 3, or (C═O)(CH₂)₁₋₅ and m is 1; wherein        W¹ is optionally interrupted with non-peroxide O, S, C═O, or        NR^(c);    -   n is 0, 1, 2, or 3; each        represents an optional double bond;    -   R³ is hydrogen, (C₁-C₁₀)alkyl, hydroxy(C₁-C₁₀)alkyl or        (C₁-C₁₀)alkoxy; and    -   R⁴ is hydroxyl (—OH), phosphate (—OPO₃H₂), phosphonate        (—CH₂PO₃H₂), or alpha-substituted phosphonate;    -   each R^(a), R^(b), R^(c), and R^(d) is independently hydrogen,        or (C₁-C₁₀)alkyl; or    -   a pharmaceutically acceptable salt or ester thereof.

-   2. The compound of embodiment 1, wherein R¹ is hydrogen, fluorine,    chlorine, bromine, trifluoro-methyl, methoxy, (C₁-C₆)alkyl,    (C₁-C₆)haloalkyl, (C₁-C₆)alkyl substituted with, alkoxy or cyano,    alkyl-substituted aryl, aryl-substituted alkyl, or aryl-substituted    arylalkyl.

-   3. The compound of embodiment 2, wherein R¹ is hydrogen,    trifluoro-methyl, or —CH₂CF₃.

-   4. The compound of embodiment 2, wherein R¹ is benzyl, phenylethyl,    or benzyl substituted with methyl.

-   5. The compound of any of embodiments 1-4, wherein R² is

-   6. The compound of embodiment 5, wherein R² is:

-   7. The compound of embodiment 6, wherein R² is

-   8. The compound of any of embodiments 1-4, wherein R² is:

-   -   wherein Z² is (CH₃)₃C—, CH₃CH₂(CH₃)₂C—, CH₃CH₂CH₂—,        CH₃(CH₂)₂CH₂—, CH₃(CH₂)₄CH₂—, (CH₃)₂CHCH₂—, (CH₃)₃CCH₂—,        CH₃CH₂O—, (CH₃)₂CHO—, or CF₃CH₂CH₂— or a group having the        formula:

-   9. The compound of embodiment 8, wherein R² is:

-   10. The compound of embodiment 9, wherein R² is:

-   11. The compound of any of embodiments 1-4, wherein R² is:

-   12. The compound of embodiment 11, wherein R² is

-   13. The compound of any of embodiments 1-4, wherein R² is    (C₁-C₂₀)alkyl, or (C₁-C₂₀)alkoxy.-   14. The compound of embodiment 13, wherein R² is (C₁-C₁₀)alkyl,    (C₂-C₁₀)alkenyl and (C₂-C₁₀)alkynyl or (C₁-C₁₀)alkoxy optionally    substituted with carbonyl (C═O) or oxime (C═NR^(d)) groups.-   15. The compound of embodiment 14, wherein R² is methyl, ethyl,    propyl, butyl, pentyl, hexyl, heptyl, octyl, trifluoromethyl,    trifluoroethyl, trifluoromethoxy, trifluoroethoxy, methoxy, ethoxy,    propoxy, butoxy, pentoxy, heptoxy, or octoxy.-   16. The compound of any of embodiments 1-15, wherein each of X¹, Y¹    and Z¹ is CH₂.-   17. The compound of any of embodiments 1-16, wherein R³ is hydrogen,    methyl, hydroxymethyl, ethyl, hydroxyethyl, propyl, or isopropyl.-   18. The compound of embodiment 17, wherein R³ is hydrogen, methyl,    hydroxymethyl, ethyl, or hydroxyethyl.-   19. The compound of any of embodiments 1-18, having the Formula

-   20. A method for prevention or treatment of a pathological condition    or symptom in a mammal, wherein the activity of sphingosine    1-phosphate receptors is implicated and agonism of such activity is    desired, comprising administering to said mammal an effective amount    of a compound of any of embodiments 1-19.-   21. The method of embodiment 19, wherein the pathological condition    is an autoimmune disease.-   22. The method of embodiment 21, wherein the autoimmune disease is    uveitis, type I diabetes, rheumatoid arthritis, inflammatory bowel    diseases, or multiple sclerosis.-   23. The method of embodiment 22, wherein the autoimmune disease is    multiple sclerosis.-   24. The method of embodiment 23, wherein the pathological condition    is altering lymphocyte trafficking.-   25. The method of embodiment 24, wherein the treatment is altering    of lymphocyte trafficking.-   26. The method of embodiment 25, wherein lymphocyte trafficking    provides prolonged allograft survival.-   27. The method of embodiment 26, wherein the allograft is for    transplantation.-   28. A method for prevention or treatment of a pathological condition    or symptom in a mammal, wherein the activity S1P lyase implicated    and inhibition of the S1P lyase is desired, comprising administering    to said mammal an effective amount of a compound of any of    embodiments 1-19.-   29. A compound of any of claims 1-19 for use in medical therapy.-   30. Use of a compound of any of embodiments 1-19, to prepare a    medicament useful for prevention or treatment of a pathological    condition or symptom in a mammal, wherein the activity of    sphingosine 1-phosphate receptors is implicated.-   31. The use of embodiment 30, wherein the medicament comprises a    carrier.-   32. The use of embodiment 31, wherein the carrier is a liquid.

Exemplary compounds of the invention have Formulas

Additional compounds of Formula IA or IB are illustrated in table 1,below.

TABLE 1

Compound R^(e) XX

XXI

XXII

XXIII

XXIV

XXV

XXXI

The compounds having formulas XX through XXV or XXXI also include allenantiomers thereof such as:

where the compounds include each of the R^(e) groups from Table 1.

In another aspect, the invention provides S1P receptor pro-drugcompounds having the general structure of Formula I, is provided by acompound with a mono-substituted tetralin ring system that has thestructure (VIII). In some embodiments of structure (I), the compound(e.g., X and XI) has only a single chiral center and that the aminocarbon is pro-chiral, i.e., will become chiral followingenzyme-catalyzed phosphorylation.

Without wishing to be bound by any particular theory, it is expectedthat the compounds described herein are pro-drugs, i.e., are activatedby phosphorylation of the primary alcohol to form themono-phosphorylated analog. Additionally, the active drugs are expectedto be agonists at the S1P type 1 receptor.

In cases where compounds are sufficiently basic or acidic to form stablenontoxic acid or base salts, preparation and administration of thecompounds as pharmaceutically salts may be appropriate. Examples ofpharmaceutically acceptable salts are organic acid addition salts formedwith acids which form a physiological acceptable anion, for example,tosylate, methanesulfonate, acetate, citrate, malonate, tartarate,succinate, benzoate, ascorbate, α-ketoglutarate, and α-glycerophosphate.Suitable inorganic salts may also be formed, including hydrochloride,sulfate, nitrate, bicarbonate, and carbonate salts.

Pharmaceutically acceptable salts may be obtained using standardprocedures well known in the art, for example by reacting a sufficientlybasic compound such as an amine with a suitable acid affording aphysiologically acceptable anion. Alkali metal (for example, sodium,potassium or lithium) or alkaline earth metal (for example calcium)salts of carboxylic acids can also be made.

Pharmaceutically-acceptable base addition salts can be prepared frominorganic and organic bases. Salts derived from inorganic bases, includeby way of example only, sodium, potassium, lithium, ammonium, calciumand magnesium salts. Salts derived from organic bases include, but arenot limited to, salts of primary, secondary and tertiary amines, such asalkyl amines, dialkyl amines, trialkyl amines, substituted alkyl amines,di(substituted alkyl) amines, tri(substituted alkyl) amines, alkenylamines, dialkenyl amines, trialkenyl amines, substituted alkenyl amines,di(substituted alkenyl) amines, tri(substituted alkenyl) amines,cycloalkyl amines, di(cycloalkyl) amines, tri(cycloalkyl) amines,substituted cycloalkyl amines, disubstituted cycloalkyl amine,trisubstituted cycloalkyl amines, cycloalkenyl amines, di(cycloalkenyl)amines, tri(cycloalkenyl) amines, substituted cycloalkenyl amines,disubstituted cycloalkenyl amine, trisubstituted cycloalkenyl amines,aryl amines, diaryl amines, triaryl amines, heteroaryl amines,diheteroaryl amines, triheteroaryl amines, heterocyclic amines,diheterocyclic amines, triheterocyclic amines, mixed di- and tri-amineswhere at least two of the substituents on the amine are different andare selected from the group consisting of alkyl, substituted alkyl,alkenyl, substituted alkenyl, cycloalkyl, substituted cycloalkyl,cycloalkenyl, substituted cycloalkenyl, aryl, heteroaryl, heterocyclic,and the like. Also included are amines where the two or threesubstituents, together with the amino nitrogen, form a heterocyclic orheteroaryl group. Examples of suitable amines include, by way of exampleonly, isopropylamine, trimethyl amine, diethyl amine, tri(iso-propyl)amine, tri(n-propyl) amine, ethanolamine, 2-dimethylaminoethanol,tromethamine, lysine, arginine, histidine, caffeine, procaine,hydrabamine, choline, betaine, ethylenediamine, glucosamine,N-alkylglucamines, theobromine, purines, piperazine, piperidine,morpholine, N-ethylpiperidine, and the like. It should also beunderstood that other carboxylic acid derivatives would be useful in thepractice of this invention, for example, carboxylic acid amides,including carboxamides, lower alkyl carboxamides, dialkyl carboxamides,and the like.

The compounds of Formula I can be formulated as pharmaceuticalcompositions and administered to a mammalian host, such as a humanpatient in a variety of forms adapted to the chosen route ofadministration, i.e., orally or parenterally, by intravenous,intramuscular, topical or subcutaneous routes.

Thus, the present compounds may be systemically administered, e.g.,orally, in combination with a pharmaceutically acceptable vehicle suchas an inert diluent or an assimilable edible carrier. They may beenclosed in hard or soft shell gelatin capsules, may be compressed intotablets, or may be incorporated directly with the food of the patient'sdiet. For oral therapeutic administration, the active compound may becombined with one or more excipients and used in the form of ingestibletablets, buccal tablets, troches, capsules, elixirs, suspensions,syrups, wafers, and the like. Such compositions and preparations shouldcontain at least about 0.1% of active compound. The percentage of thecompositions and preparations may, of course, be varied and mayconveniently be between about 2 to about 60% of the weight of a givenunit dosage form. The amount of active compound in such therapeuticallyuseful compositions is such that an effective dosage level will beobtained.

The tablets, troches, pills, capsules, and the like may also contain thefollowing: binders such as gum tragacanth, acacia, corn starch orgelatin; excipients such as dicalcium phosphate; a disintegrating agentsuch as corn starch, potato starch, alginic acid and the like; alubricant such as magnesium stearate; and a sweetening agent such assucrose, fructose, lactose or aspartame or a flavoring agent such aspeppermint, oil of wintergreen, or cherry flavoring may be added. Whenthe unit dosage form is a capsule, it may contain, in addition tomaterials of the above type, a liquid carrier, such as a vegetable oilor a polyethylene glycol. Various other materials may be present ascoatings or to otherwise modify the physical form of the solid unitdosage form. For instance, tablets, pills, or capsules may be coatedwith gelatin, wax, shellac or sugar and the like. A syrup or elixir maycontain the active compound, sucrose or fructose as a sweetening agent,methyl and propylparabens as preservatives, a dye and flavoring such ascherry or orange flavor. Of course, any material used in preparing anyunit dosage form should be pharmaceutically acceptable and substantiallynon-toxic in the amounts employed. In addition, the active compound maybe incorporated into sustained-release preparations and devices.

The active compound may also be administered intravenously orintraperitoneally by infusion or injection. Solutions of the activecompound or its salts can be prepared in water, optionally mixed with anontoxic surfactant. Dispersions can also be prepared in glycerol,liquid polyethylene glycols, triacetin, and mixtures thereof and inoils. Under ordinary conditions of storage and use, these preparationscontain a preservative to prevent the growth of microorganisms.

The pharmaceutical dosage forms suitable for injection or infusion caninclude sterile aqueous solutions or dispersions or sterile powderscomprising the active ingredient which are adapted for theextemporaneous preparation of sterile injectable or infusible solutionsor dispersions, optionally encapsulated in liposomes. In all cases, theultimate dosage form should be sterile, fluid and stable under theconditions of manufacture and storage. The liquid carrier or vehicle canbe a solvent or liquid dispersion medium comprising, for example, water,ethanol, a polyol (for example, glycerol, propylene glycol, liquidpolyethylene glycols, and the like), vegetable oils, nontoxic glycerylesters, and suitable mixtures thereof. The proper fluidity can bemaintained, for example, by the formation of liposomes, by themaintenance of the required particle size in the case of dispersions orby the use of surfactants. The prevention of the action ofmicroorganisms can be brought about by various antibacterial andantifungal agents, for example, parabens, chlorobutanol, phenol, sorbicacid, thimerosal, and the like. In many cases, it will be preferable toinclude isotonic agents, for example, sugars, buffers or sodiumchloride. Prolonged absorption of the injectable compositions can bebrought about by the use in the compositions of agents delayingabsorption, for example, aluminum monostearate and gelatin.

Sterile injectable solutions are prepared by incorporating the activecompound in the required amount in the appropriate solvent with variousof the other ingredients enumerated above, as required, followed byfilter sterilization. In the case of sterile powders for the preparationof sterile injectable solutions, the preferred methods of preparationare vacuum drying and the freeze drying techniques, which yield a powderof the active ingredient plus any additional desired ingredient presentin the previously sterile-filtered solutions.

For topical administration, the present compounds may be applied in pureform, i.e., when they are liquids. However, it will generally bedesirable to administer them to the skin as compositions orFormulations, in combination with a dermatologically acceptable carrier,which may be a solid or a liquid.

Suitable solid carriers include finely divided solids such as talc,clay, microcrystalline cellulose, silica, alumina and the like. Usefulliquid carriers include water, alcohols or glycols orwater-alcohol/glycol blends, in which the present compounds can bedissolved or dispersed at effective levels, optionally with the aid ofnon-toxic surfactants. Adjuvants such as fragrances and additionalantimicrobial agents can be added to optimize the properties for a givenuse. The resultant liquid compositions can be applied from absorbentpads, used to impregnate bandages and other dressings, or sprayed ontothe affected area using pump-type or aerosol sprayers.

Thickeners such as synthetic polymers, fatty acids, fatty acid salts andesters, fatty alcohols, modified celluloses or modified mineralmaterials can also be employed with liquid carriers to form spreadablepastes, gels, ointments, soaps, and the like, for application directlyto the skin of the user.

Examples of useful dermatological compositions which can be used todeliver the compounds of Formula I to the skin are known to the art; forexample, see Jacquet et al. (U.S. Pat. No. 4,608,392), Geria (U.S. Pat.No. 4,992,478), Smith et al. (U.S. Pat. No. 4,559,157) and Wortzman(U.S. Pat. No. 4,820,508).

Useful dosages of the compounds of Formula I can be determined bycomparing their in vitro activity, and in vivo activity in animalmodels. Methods for the extrapolation of effective dosages in mice, andother animals, to humans are known to the art; for example, see U.S.Pat. No. 4,938,949.

Generally, the concentration of the compound(s) of Formulas I-IV in aliquid composition, such as a lotion, will be from about 0.1-25 wt-%,preferably from about 0.5-10 wt-%. The concentration in a semi-solid orsolid composition such as a gel or a powder will be about 0.1-5 wt-%,preferably about 0.5-2.5 wt-%.

The amount of the compound, or an active salt or derivative thereof,required for use in treatment will vary not only with the particularsalt selected but also with the route of administration, the nature ofthe condition being treated and the age and condition of the patient andwill be ultimately at the discretion of the attendant physician orclinician.

In general, however, a suitable dose will be in the range of from about0.5 to about 100 mg/kg, e.g., from about 10 to about 75 mg/kg of bodyweight per day, such as 3 to about 50 mg per kilogram body weight of therecipient per day, preferably in the range of 6 to 90 mg/kg/day, mostpreferably in the range of 15 to 60 mg/kg/day.

The compound is conveniently administered in unit dosage form; forexample, containing 5 to 1000 mg, conveniently 10 to 750 mg, mostconveniently, 50 to 500 mg of active ingredient per unit dosage form.

Ideally, the active ingredient should be administered to achieve peakplasma concentrations of the active compound of from about 0.5 to about75 μM, preferably, about 1 to 50 μM, most preferably, about 2 to about30 μM. This may be achieved, for example, by the intravenous injectionof a 0.05 to 5% solution of the active ingredient, optionally in saline,or orally administered as a bolus containing about 1-100 mg of theactive ingredient. Desirable blood levels may be maintained bycontinuous infusion to provide about 0.01-5.0 mg/kg/hr or byintermittent infusions containing about 0.4-15 mg/kg of the activeingredient(s).

The desired dose may conveniently be presented in a single dose or asdivided doses administered at appropriate intervals, for example, astwo, three, four, or more sub-doses per day. The sub-dose itself may befurther divided, e.g., into a number of discrete loosely spacedadministrations; such as multiple inhalations from an insufflator or byapplication of a plurality of drops into the eye.

The method of the invention includes a kit comprising an inhibitoridentified in the invention and an instructional material whichdescribes administering the inhibitor or a composition comprising theinhibitor to a cell or a subject. This should be construed to includeother embodiments of kits that are known to those skilled in the art,such as a kit comprising a (preferably sterile) solvent suitable fordissolving or suspending the composition of the invention prior toadministering the compound to a cell or a subject. Preferably, thesubject is a human.

The term “instructional material” includes a publication, a recording, adiagram, or any other medium of expression which can be used tocommunicate the usefulness of the composition of the invention for itsdesignated use. The instructional material of the kit of the inventionmay, for example, be affixed to a container which contains thecomposition or be shipped together with a container which contains thecomposition. Alternatively, the instructional material may be shippedseparately from the container with the intention that the instructionalmaterial and the composition be used cooperatively by the recipient.

In accordance with the present invention, as described above or asdiscussed in the Examples below, there can be employed conventionalchemical, cellular, histochemical, biochemical, molecular biology,microbiology, and in vivo techniques which are known to those of skillin the art. Such techniques are explained fully in the literature.

Without further description, it is believed that one of ordinary skillin the art can, using the preceding description and the followingillustrative examples, make and utilize the compounds of the presentinvention.

Processes for preparing compounds of Formula I or for preparingintermediates useful for preparing compounds of Formula I are providedas further embodiments of the invention. Intermediates useful forpreparing compounds of Formula I are also provided as furtherembodiments of the invention. The processes are provided as furtherembodiments of the invention and are illustrated in the schemes hereinwherein the meanings of the generic radicals are as given above unlessotherwise qualified.

Processes for preparing compounds of Formula I or for preparingintermediates useful for preparing compounds of Formula I are providedas further embodiments of the invention. Intermediates useful forpreparing compounds of Formula I are also provided as furtherembodiments of the invention. The compounds of the invention can beprepared using starting materials and methods known in the art.

The syntheses of target molecules are illustrated in Scheme 1, Scheme 2,and Scheme 3 (See FIGS. 3-5).

The invention is now described with reference to the following Examplesand Embodiments. Without further description, it is believed that one ofordinary skill in the art can, using the preceding description and thefollowing illustrative examples, make and utilize the disclosedcompounds. The following working examples therefore, are provided forthe purpose of illustration only, point out the preferred embodiments,and are not to be construed as limiting in any way the remainder of thedisclosure. Therefore, the examples should be construed to encompass anyand all variations which become evident as a result of the teachingprovided herein.

Scheme 1 Example 1 Compound 2

6-Hydroxy-2-tetralone (4.0 g, 24.7 mmol) is dissolved in methylenechloride (50 mL) and cooled to 0° C. under inert atmosphere.Trifluoromethyl sulfonyl anhydride (5.55 g, 25.0 mmol) is added to thereaction. 2,6-Lutidine (3.2 g, 30 mmol) is introduced dropwise over 10minutes and the reaction is stirred at 0° C. for an additional 2 hours.The reaction volume is reduced to about 10 mL in vacuo and filtered toremove insoluble material. The filter cake is washed with ether (2×25mL), the organic layers combined and solvent removed in vacuo leaving athick residue that is utilized in the next step without furtherpurification.

Example 2 Compound 3

An octyl boronate intermediate is first generated by adding 1-octene(3.95 g, 36.0 mmol) to a solution of 9-BBN in THF (80.6 mL of a 0.5 Msolution, 40.3 mmol). The reaction is stirred at room temperatureovernight, then an aqueous solution of sodium hydroxide (7.0 mL of a 3.0M solution) is added, followed by the addition of the crude residue(compound 2) from Example 1 dissolved in THF (25 mL) and Pd[P(Ph)₃]₄(0.900 g, 0.785 mmol). The reaction is brought to reflux and maintainedfor 12 hours under an inert atmosphere. The reaction is then cooled,filtered, concentrated in vacuo and the residue purified using silicagel chromatography with hexanes/ethyl acetate (75:25) as solvent.Purified 6-octyl-2-tetralone is a clear liquid (4.13 g, 16.1 mmol, 63%).

Example 3 Compound 3 (Alternate Route)

An octyl boronate intermediate is first generated by adding 1-octene(1.38 g, 14.1 mmol) to a solution of 9-BBN in THF (28.3 mL of a 0.5 Msolution, 14.1 mmol). The reaction is stirred at room temperatureovernight, then an aqueous solution of sodium hydroxide (5.0 mL of a 3.0M solution) is added, followed by the addition of 6-bromo-2-tetralone(2.40 g, 10.0 mmol) and Pd[P(Ph)₃]₄ (0.310 g, 0.270 mmol). The reactionis brought to reflux and maintained for 12 hours under an inertatmosphere. The reaction is then cooled, filtered, concentrated in vacuoand the residue purified using silica gel chromatography withhexanes/ethyl acetate (75:25) as solvent to provide 6-octyl-2-tetralone(2.35 g, 8.5 mmol, 85%) as a clear liquid.

Example 4 Compound 4

Potassium hydride (1.07 g of a 30% w/w suspension in mineral oil, 8.0mmol) is washed several times with pentane to remove the mineral oil,then THF (25 mL) is added and the mixture cooled to −40° C.6-Octyl-2-tetralone (1.95 g, 7.0 mmol) dissolved in THF (20 mL) is addeddropwise over 30 minutes at −40° C. The reaction is allowed to warm to0° C. for 30 minutes, then returned to −40° C.2,2-Dimethyl-5-formyl-1,3-dioxan-5-yl)acetamide (1.65 g, 8.2 mmol) inTHF (20 mL) is added dropwise and the reaction maintained at −40° C. for2 hours. The reaction is quenched by the addition of acetic anhydride(1.02 g, 10 mmol) at −40° C. followed by warming the reaction to roomtemperature and stirring for 30 minutes. P-toluenesulfonic acid (˜10 mg)is added and the reaction brought to reflux and maintained for 1 hour.The reaction is cooled partitioned between ether (200 mL) and aqueoussodium bicarbonate (5% w/w, 100 mL). The organic layer is removed,washed with aqueous bicarbonate (1×50 mL), then dried and the solventsremoved in vacuo. The residue is purified using silica gelchromatography with hexanes (60%)-ethyl acetate (40%) to afford compound4.

Example 5 Compound 5

Compound 4 (658 mg, 1.42 mmol) is dissolved in ethanol (15 mL).Palladium on activated carbon (90 mg of 10 wt. %) is added, the mixtureis cooled to 0° C. and a hydrogen atmosphere is introduced. The reactionis stirred vigorously overnight at 0° C., then filtered through Celiteand the solvent removed in vacuo. Compound 5 is purified usingchromatography on silica gel using hexanes (60%)-ethyl acetate (40%).

Example 6 Compound 6

Compound 5 (298 mg, 0.64 mmol) is dissolved in ethanol (10 mL) andsodium borohydride (10 mg) is added. The reaction mixture is stirred atroom temperature for 3 hours. The mixture is partitioned between ether(30 mL) and water (10 mL). The organic layer is washed once with water(1×10 mL), then brine and dried over MgSO₄. The combined organic layersare removed in vacuo and the residue, containing the product isomericalcohols (compound 6), is used in the next step without purification.

Example 7 Compound 7

The crude compound 6 is dissolved in toluene (10 mL) containingp-toluenesulfonic acid (˜5 mg) containing activated 3A molecular sievespheres (25 mg). The reaction is heated to 70° C. under an inertatmosphere for 4 hours, cooled, filtered and partitioned between ether(25 mL) and aqueous sodium bicarbonate (5% w/w, 10 mL). The organiclayer is washed once with aqueous bicarbonate solution (10 mL), thenwith brine and dried over MgSO₄. The solvents are removed in vauco andthe residue is purified by chromatography on silica gel using hexanes(50%)-ethyl acetate (50%).

Example 8 Compound 8

Compound 7 (138 mg, 0.29 mmol) is dissolved in ethanol (10 mL) andpalladium on activated carbon (30 mg of 10 wt. %) is added. The reactionis maintained at room temperature, and a hydrogen atmosphere isintroduced. The reaction is stirred vigorously overnight and thenfiltered through Celite. Aqueous hydrochloric acid (1.0 mL of a 5 Nsolution) is introduced and the reaction refluxed for 3 hours. Thereaction is partitioned between ether (30 mL) and aqueous sodiumbicarbonate solution (5% w/w, 10 mL). The organic layer is washed withaqueous bicarbonate (1×10 mL), then with brine and dried over MgSO₄. Thesolvent removed in vacuo. Compound 8 is purified using chromatography onsilica gel using hexanes (30%)-ethyl acetate (70%).

Example 9 Compound 9

Compound 8 (93 mg, 0.23 mmol) is dissolved in methanol (3.0 mL) and LiOH(100 mg, 1.54 mmol) is added. The reaction is heated at reflux for 2hours, then partitioned between methylene chloride (10 mL) and water (5mL). The aqueous layer is extracted with methylene chloride (2×5 mL) andthe combined organics are then dried over MgSO₄. The organic layer isevaporated to dryness. The residue is purified using silica gelchromatography with methanol (25%)-methylene chloride (75%) to furnishcompound 9.

Example 10 Compound 10

Compound 7 (See: FIG. 3, Scheme 1) (100 mg, 0.21 mmol) is dissolved inethanol (10 mL), aqueous hydrochloric acid (1.0 mL of a 5 N solution) isintroduced and the reaction refluxed for 6 hours. The reaction iscooled, then partitioned between ether (30 mL) and aqueous sodiumbicarbonate solution (5% w/w, 10 mL). The organic layer is separated,then washed with aqueous bicarbonate (1×10 mL), and brine, dried overMgSO₄, and the solvent removed in vacuo. Compound 10 is provided withchromatography on silica gel using hexanes (30%)-ethyl acetate (70%).

Example 11 Compound 11

Compound 10 (69 mg, 0.17 mmol) is dissolved in methanol (3.0 mL) andLiOH (100 mg, 1.54 mmol) is added. The reaction is heated at reflux for2 hours, partitioned between methylene chloride (10 mL) and water (5mL). The aqueous layer is extracted with methylene chloride (2×5 mL) andthe combined organics are then dried over MgSO₄. The organic layer isevaporated to dryness. The residue is purified using silica gelchromatography with methanol (25%)-methylene chloride (75%) to givecompound 11.

The assays below are standard literature reported assays known in theart for confirming and quantifying the activity of the disclosedcompounds.

Example 12 Sphingosine Kinase Assay

Recombinant sphingosine kinase type 2 (SPHK2) is prepared by forcing theexpression of the mouse or human recombinant enzyme by transfecting therelevant plasmid DNA into HEK293T or CHO K1 cells. After about 60 hours,cells are harvested, broken and the non-microsomal (e.g., soluble)fraction is retained. The broken cell supernatant fluid containing therecombinant enzyme is mixed with test compounds (e.g., FTY720, AA151,VIII and XVIII) (5-50 micromolar) and γ-32P-ATP and incubated for0.5-2.0 hours at 37° C. The lipids in the reaction mixture are extractedinto an organic solvent and displayed by normal phase thin layerchromatography. The radio-labeled bands are detected by autoradiography,scraped from the plate and quantified by scintillation counting. Thetest compounds are used at a concentration of about 50 μM, incubationtime is about 20 minutes.

Example 13 GTPγS-35 Binding Assay

This assay illustrates agonist activation of G protein coupled receptors(GPCRs) in isolation. The assay forces expression concomitantly of arecombinant GPCR (e.g., the S1P1-5 receptor) and each of the threesubunits (e.g., α-i2, β-1, and γ-2) of a heterotrimeric G protein in aHEK293T cell by transfecting the cell with four plasmid DNAs encodingthe respective proteins. About 60 hours after transfection cells areharvested, broken, the nucleus discarded, and the crude microsomes areprepared from the remainder. Agonist (e.g., S1P) stimulation of thereceptor-G protein complex on the microsomes results in the exchange ofGTP for GDP on the α-subunit in a dose-dependent manner. The GTP-boundα-subunit is detected using a GTP analog (GTPγS-35), which is aradionuclide (sulfur-35) labeled phosphothionate that is not hydrolyzedto GDP. The microsomes with the adherent G proteins are collected byfiltration and the bound GTPγS-35 quantified in a liquid scintillationcounter. The assay provides relative potency (EC₅₀ values) and maximumeffect (efficacy, E_(max)). Antagonist activity is detected as rightwardshifts in the agonist dose-response curve in the presence of a fixedamount of antagonist. If the antagonist behaves competitively, theaffinity of the receptor/antagonist pair (K_(i)) can be determined. Theassay is described in Davis, M. D., J. J. Clemens, T. L. Macdonald andK. R. Lynch (2005) “S1P Analogs as Receptor Antagonists” Journal ofBiological Chemistry, vol. 280, pp. 9833-9841.

Example 14 Lymphopenia Assay

The test compounds (e.g., primary alcohols) are dissolved in 2%hydroxypropyl beta-cyclodextrin and introduced into groups of mice byoral gavage at doses from 0.01, 1.0 and 10 mg/kg body weight. Atintervals, e.g., 24 hours, 48 hours, or 96 hours the mice are lightlyanesthetized and ca. 0.1 mL of blood is drawn from the orbital sinus.The number of lymphocytes (in thousands per microliter of blood; normalis 4-11) is determined using a Hemavet blood analyzer.

Example 15 Heart Rate Assay

Mice are dosed with test compounds (intravenous, 3 mg/kg) or vehicle (2%hydroxypropyl beta-cyclodextrin) and the heart rate measured at 1 hourpost dosing. Heart rate is captured in unrestrained, conscious animalsusing the ECGenie™ system.

The abbreviations used herein have their conventional meaning within theclinical, chemical, and biological arts. In the case of anyinconsistencies, the present disclosure, including any definitionstherein will prevail.

The disclosures of each and every patent, patent application, andpublication cited herein are expressly incorporated herein by referencein their entirety into this disclosure. Illustrative embodiments of thisdisclosure are discussed and reference has been made to possiblevariations within the scope of this disclosure. These and othervariations and modifications in the disclosure will be apparent to thoseskilled in the art without departing from the scope of the disclosure,and it should be understood that this disclosure and the claims shownbelow are not limited to the illustrative embodiments set forth herein.

1. A compound of Formula I:

wherein X¹, Y¹ and Z¹ are independently O, CR^(a), CR^(a)R^(b), N, NR^(c), or S; R¹ and R² are independently hydrogen, halo, halo(C₁-C₁₀)alkyl, cyano, —NR^(a)R^(b), (C₁-C₂₀)alkyl, (C₂-C₂₀)alkenyl, (C₂-C₂₀)alkynyl, (C₁-C₂₀)alkoxy, (C₂-C₂₆)alkoxyalkyl, (C₃-C₁₂)cycloalkyl, (C₆-C₁₀)aryl, (C₇-C₃₀)arylalkyl, (C₂-C₁₀)heterocyclic, (C₄-C₁₀)heteroaryl, or (C₄-C₁₀)heteroaryl(C₁-C₂₀)alkyl; wherein the alkyl, alkenyl, alkynyl, cycloalkyl, aryl, heterocyclic, or heteroaryl groups of R¹ and R² independently are optionally perfluorinated or optionally substituted with 1, 2, 3, or 4 groups where the substituent groups are independently hydroxy, halo, cyano, (C₁-C₁₀)alkoxy, C₆-aryl, (C₇-C₂₄)arylalkyl, oxo (═O), or imino (═NR^(d)), wherein one or more of the carbon atoms in the R¹ or R² alkyl groups can be independently replaced with non-peroxide oxygen, sulfur or NR^(c); the alkyl groups of R³ are optionally substituted with 1, or 2 hydroxy groups; or R² can be a group having formula II, III, IV, V, or VI;

wherein each R⁷, R⁸, R⁹, R¹⁰, R¹¹, R¹², R¹³, and R¹⁴ is independently O, S, C, CR¹⁵, CR¹⁶R¹⁷, C═O, N or NR¹⁸; each R¹⁵, R¹⁶ and R¹⁷ is independently hydrogen, halo, (C₁-C₁₀)alkyl, (C₆-C₁₀)aryl, (C₁-C₁₀)alkyl substituted with halo, hydroxy, (C₁-C₁₀)alkoxy, or cyano; and where R¹⁸ can be hydrogen or (C₁-C₁₀)alkyl; wherein Z² is hydrogen, halo, halo(C₁-C₁₀)alkyl, cyano, —NR^(a)R^(b), (C₁-C₂₀)alkyl, (C₂-C₂₀)alkenyl, (C₂-C₂₀)alkynyl, (C₁-C₂₀)alkoxy, (C₂-C₂₆)alkoxyalkyl, (C₃-C₁₂)cycloalkyl, (C₆-C₁₀)aryl, (C₇-C₃₀)arylalkyl, (C₂-C₁₀)heterocyclic, (C₄-C₁₀)heteroaryl, or (C₄-C₁₀)heteroaryl(C₁-C₂₀)alkyl; wherein the alkyl, alkenyl, alkynyl, cycloalkyl, aryl, heterocyclic, or heteroaryl groups of Z² are optionally perfluorinated or optionally substituted with 1, 2, 3, or 4 groups where the substituent groups are independently hydroxy, halo, cyano, (C₁-C₁₀)alkoxy, C₆₋aryl, (C₇-C₂₄)arylalkyl, oxo (═O), or imino (═NR^(d)), wherein one or more of the carbon atoms in the Z² alkyl groups can be independently replaced with non-peroxide oxygen, sulfur or NR^(c);

indicates one or more optional double bonds; wherein Y² is a bond, O, S, C═O, or NR^(c), CH₂; W¹ is a bond, —CH₂— and m is 1, 2, or 3, or W¹ is (C═O)(CH₂)₁₋₅ and m is 1; wherein W¹ is optionally interrupted with non-peroxide O, S, C═O, or NR^(c); n is 0, 1, 2, or 3; one

 represents an optional double bond; R³ is hydrogen, (C₁-C₁₀)alkyl, hydroxy(C₁-C₁₀)alkyl or (C₁-C₁₀)alkoxy; and R⁴ is hydroxyl (—OH), phosphate (—OPO₃H₂), phosphonate (—CH₂PO₃H₂), or alpha-substituted phosphonate; each R^(a), R^(b), R^(c), and R^(d) is independently hydrogen, CF₃ or (C₁-C₁₀)alkyl; or a pharmaceutically acceptable salt or ester thereof.
 2. The compound of claim 1, wherein R¹ is hydrogen, fluorine, chlorine, bromine, trifluoro-methyl, methoxy, (C₁-C₆)alkyl, (C₁-C₆)haloalkyl, (C₁-C₆)alkyl substituted with, alkoxy or cyano, alkyl-substituted aryl, aryl-substituted alkyl, or aryl-substituted arylalkyl.
 3. The compound of claim 2, wherein R¹ is hydrogen, trifluoro-methyl, or —CH₂CF₃.
 4. The compound of claim 2, wherein R¹ is benzyl, phenylethyl, or benzyl substituted with methyl.
 5. The compound of claim 1, wherein R² is


6. The compound of claim 1, wherein R² is:

wherein Z² is (CH₃)₃C—, CH₃CH₂(CH₃)₂C—, CH₃CH₂CH₂—, CH₃(CH₂)₂CH₂—, CH₃(CH₂)₄CH₂—, (CH₃)₂CHCH₂—, (CH₃)₃CCH₂—, CH₃CH₂O—, (CH₃)₂CHO—, or CF₃CH₂CH₂— or a group having the formula:


7. The compound of claim 6, wherein R² is:


8. The compound of claim 1, wherein R² is:


9. The compound of claim 8, wherein R² is


10. The compound of claim 1, wherein R² is methyl, ethyl, propyl, butyl, pentyl, hexyl, heptyl, octyl, trifluoromethyl, trifluoroethyl, trifluoromethoxy, trifluoroethoxy, methoxy, ethoxy, propoxy, butoxy, pentoxy, heptoxy, or octoxy.
 11. The compound of claim 1, wherein X¹, Y¹ and Z¹ are CH₂.
 12. The compound of claim 1, wherein R³ is hydrogen, methyl, hydroxymethyl, ethyl, or hydroxyethyl.
 13. The compound of claim 1, having the Formula


14. A method for treatment of a pathological condition or symptom in a mammal, wherein the pathological condition is an autoimmune disease wherein the autoimmune disease is uveitis, type I diabetes, rheumatoid arthritis, inflammatory bowel diseases, or multiple sclerosis, comprising administering to said mammal an effective amount of a compound claim
 1. 15. The method of claim 14, wherein the autoimmune disease is multiple sclerosis.
 16. The method of claim 14, wherein the pathological condition or symptom is altering of lymphocyte trafficking.
 17. The method of claim 16, wherein altering lymphocyte trafficking provides prolonged allograft survival. 